Objective: In this study, in order to compare PCR andcultivation methods to determine the incidence ofinfections due to  M. hominis  and  M. fermentans  inwomen genitourinary tract  100 genital swabs and 100urine samples obtained from women with genitourinarytract (GUT) infection were studied.Method:  Genital swab and urine samples wereinoculated and transported with a selectivemycoplasma  transport media. After incubation at 37°Cfor 18-24 hours 0.3 mL medium samples weretransferred to the specific solid medium formycoplasma.  The agar plates were incubated at thesame atmosphere conditions (5% CO2 and 95% N2 at37°) for 48-72 hours. Characteristic mycoplasmacolonies were determined by staining with Dienneísstain and examined by x10 microscope objective. Thegenital swab and urine samples were also analyzed bya nested PCR protocol with genus specific MCGpF11,R23-TR, R16-2 and MCGR21 primers. Another PCRprotocol was also performed in order to confirm thesamples which have compatible target sequences forM. fermentans by using RW004-RW005 primers. On theother hand, all other mycoplasma positive ampliconswere also digested with  VspI    in order to determinetwo DNA fragments (123bp and 113bp) which werecompatible for M. hominis in tested samples.Results: Mycoplasma    strains were isolated from 26(26%) genital swabs and 11 (11%) urine samples byusing a selective mycoplasma isolation media. Totally40 samples were found to be positive for mycoplasmaswhich consisted of target genomic sequences of  M.hominis  and  M. fermentans  in 37(37%) and 3(3%)samples respectively.Conclusions:  We  found that there could be anassociation with  M. hominis  (37%) and women withgenital infection, also with  M. fermentans  (3%) andalthough the high specificity (100%) of cultivation, ithas a low sensitivity (70.3%) and time consuming whencompared with PCR . On the other hand, we concludedthat, PCR is a sensitive and easily applicable protocolwhen genus specific primers are used for the diagnosisof mycoplasmas.Key words:  Mycoplasma hominis, mycoplasmafermentans, PCR, DNA, RFLP